【作者】 马良；

【导师】 崔同； 陈冠华；

【作者基本信息】 河北农业大学 ， 农产品加工及贮藏工程， 2004， 硕士

【摘要】 中国不仅是东方梨的发源中心，同时也是世界第一产梨大国，在漫长的栽培和食用过程中，古人已经发现了梨的许多食疗和保健作用，然而到目前为止，关于梨果实中生理活性成分方面有价值的研究报道十分罕见。在前期研究中，本研究组曾首次发现东方梨果实中有高含量的熊果苷、绿原酸等酚类活性物质，本实验研究表明熊果苷和绿原酸应该是梨果实最具代表性的活性成分。 本文利用国产高效液相色谱仪建立了测定梨果实中主要活性成分的等度高效液相色谱法和梯度淋洗高效液相色谱法，等度淋洗高效液相色谱分析方法使用HYPERSIL C₁₈ BDS色谱柱（4.6mm×200mm，5μm），流动相为甲醇：水（25∶75，v／v），磷酸调节pH值为3.20，流速0.6 ml／min，室温条件，紫外检测器波长280nm，分析时间13分。熊果苷在144-5000ng，绿原酸在200-5000ng的范围线性相关系数均为0.99998，最低检出限分别为0.153ng和0.150ng，加标回收率分别为101％和97％。对梨果实中熊果苷、绿原酸、儿茶素、表儿茶素、咖啡酸、香豆酸、槲皮素等多酚类活性成分建立了双泵高压梯度淋洗的高效液相色谱分析方法。梯度淋洗程序（min／B％）为：0／2，3／2，4／25，11／50，16／80，26／80，33／2，45／2，流动相用磷酸调pH3.20，280nm检测波长下，30分钟内样品组分全部沈出，45分钟时色谱柱重新稳定，可以分析下一个样品。采用建立的高效液相色谱法对鸭梨的花期和果实生长期活性成分含量进行了动态分析，花期的不同器官中两种成分含量均处于较高的水平，尤其熊果苷，在花芽和叶芽中含量高达1.3％左右，绿原酸为3000μg／g左右。在果实生长期，幼果中两种酚类成分含量最高，分别为1.0％和0.38％，之后快速下降，果实成熟时这些酚类成分含量降低到400μg／g以下。对东方梨系统的白梨、砂梨、秋子梨和新疆梨以及西洋梨5个种的22个品种进行了分析，东方梨果实中熊果苷的含量（17个品种平均164μg／g）高于西洋梨（5个品种平均83μg／g），其中主栽品种白梨中的鸭梨含量最高，达到400μg／g。西洋梨中的绿原酸含量更为突出，平均309μg／g，高于东方梨的含量（平均158μg／g）。这些成分在果实中的分布差别很大，熊果苷在果皮中含量最高，分别为果心和果肉的3-5倍和10-40倍。东方梨中绿原酸的分布果心含量高于果皮，而西洋梨则相反。更多还原

【Abstract】 China is not only the cradle center of eastern pear but also the biggest county in pear’s output of the world. During the lengthy process of growth and eating, some therapy function through biting and supping and many hygienically the ancients founded function. But the infrequent investigations and reports about the constituents with physiologic activity in pear fruit were founded by far. Our research team founded the upper level content of the constituents with physiologic activity for example arbutin and chlorogenic acid in eastern pear in initial stages. It was founded that Arbutin and chlorogenic acid should be the representative components with physiologic activity in eastern pear in fact by our test.Isocratic elution reversed-phase high-performance liquid chromatographic (HPLC) method and gradient elution reversed-phase high-performance liquid chromatographic (HPLC) method for the analysis of the main constituents with physiologic activity in pear fruit were developed with high-performance liquid chromatographic instrument which was made in china. Isocratic elution method make up of a reversed-phase column (4.6mmx200mm) filled with 5um HYPERSIL BDS as the stationary phase .The mobile phase was made of CH3OH-H2O (25: 75, v/v) dualistic system at pH3.20, which adjusted, by phosphoric acid. The flow rate of mobile phase was 0.6 ml / min. During 13 minutes the analysis was ended at 280nm wavelength with ultraviolet detector in room temperature. The linearity correlative coefficient are 0.99998 both arbutin and chlorogenic acid. The linear range is at 144-5000ng to arbutin and is at 200-5000ng to chlorogenic acid. The two components’ lowest detected limit was 0.153 ng and 0.150 ng respectively. Gradient elution reversed-phase HPLC method using two pumps established to the main polyphenolic compounds for example arbutin, chlorogenic acid, catechin, epicatechin, caffeic acid, p-coumaric acid, quercetin, et, al in pear fruit. Gradient elution program (min/B%): 0/2, 3/2, 4/25, 11/50, 16/80, 26/80, 33/2, 45/2 .The mobile phase at pH3.20 adjusted by phosphoric acid and detected at 280nm wavelength. In this HPLC system, the main active components were separated within 30 minutes and chromatography column condition was renewably stabilized within 45 minutes and the next sample can be run. Dynamic changes in the main constituents with physiologic activity during the florescence and the fruits development stage of "Ya" Pear were monitored by the developed HPLC methods.The contents of the two constituents in different organs during the florescence areon upper level especially the content of arbutin .The contents of arbutin reached about 1.3% in floral bud and foliar bud while the contents of chlorogenic acid obtained about 3000u.g/g .The contents of the two polyphenolic compounds in fruit let stage during the process of fruit growing are the tiptop that reached 1.0% and 0.38% respectively . Then the content of the polyphenolic compounds decline swiftly and the content of the polyphenolic compounds reduce below 400ug/g in the maturation period of pear fruit. Twenty-two varieties include P. bretschnrideri Reh., P. pyrifolia Burm Nakai, P. ussuriensis Maxim., P. sinkiangensis Yu. of eastern pear system and five varieties of Pyrus communis L. was compared and analyzed. The content of arbutin in eastern pear fruit (the average content is 164ug/g of 17 varieties) were upper than the Pyrus communis L. (the average content is 83ug/g). Ya pear is one variety of P.bretschnrideri Reh. Which mostly planting variety in eastern pear system and its content of arbutin were uppest reaching 400(4ug/g. The content of chlorogenic acid was given prominence to Pyrus communis L. with 309(ug/g average and upper than the content of eastern pear (158ug/g). The content distribution of the components was prodigiously different in pear fruit. The tiptop content of arbutin was distributing in the fruit peel and obtained 3-5 times to fruit core and 10-40 times to fruit pulp. The content distribution of chlorogenic acid fruit core was upper than frui更多还原